Antifungal 4-substituted 5,6-dihydro-4H-pyrrolo [1,2-a][1,4]benzodiazepines

ABSTRACT

The present invention concerns compounds for use as a medicine having the formula 
                         
the N-oxide forms, the salts, the quaternary amines and stereochemically isomeric forms thereof, wherein
     R 1  represents hydrogen, C 1-6 alkyl, C 1-4 alkylthio, C 1-4 alkyloxy, or halo;   R 2  represents hydrogen or C 1-6 alkyl;   R 3  represents phenyl substituted with halo, cyano, C 1-4 alkyloxy, C 1-4 alkylthio, C 1-6 alkyl, haloC 1-6 alkyl; 2-thienyl; or 3-thienyl,   R 4  represents hydrogen; or R 2  and R 4  form an extra bond,
 
which are active against dermatophytes, and their preparation; it further relates to compositions comprising them, as well as their use as a medicine.

The present application is a continuation of prior application U.S. Ser.No. 10/399,596, now U.S. Pat. No. 7,202,240, which was filed on Feb. 4,2004 as the National Stage application under 35 U.S.C. 371 ofApplication No. PCT/EP01/12287 filed Oct. 17, 2001, which claimspriority from EP 00203726.5, filed on Oct. 23, 2000, the contents ofwhich are incorporated herein by reference.

The present invention is concerned with a new class of antifungalsactive mainly against dermatophytes, and their preparation; it furtherrelates to compositions comprising them, as well as their use as amedicine.

5,6-Dihydro-4H-pyrrolo[1,2-a][1,4]benzodiazepines have been described inJ. Chem. Soc. (C), 2732-2734 (1971); J. Heterocyclic Chem., 13, 711-716(1976); and J. Heterocyclic Chem., 16, 241-244 (1979); no biologicalactivities were reported in any of these references.

The present invention concerns compounds for use as a medicine havingthe formula

the N-oxide forms, the salts, the quaternary amines and stereochemicallyisomeric forms thereof, wherein

-   R¹ represents hydrogen, C₁₋₆alkyl, C₁₋₄alkylthio, C₁₋₄alkyloxy, or    halo;-   R² represents hydrogen or C₁₋₆alkyl;-   R³ represents phenyl substituted with halo, cyano, C₁₋₄alkyloxy,    C₁₋₄alkylthio, C₁₋₆alkyl, haloC₁₋₆alkyl; 2-thienyl; or 3-thienyl;    and-   R⁴ is hydrogen; or    -   R² and R⁴ form an extra bond.

The compounds of formula (I) are novel provided that R³ is not4-methylphenyl, 4-methoxyphenyl or 4-isopropylphenyl when R¹, R² and R⁴are hydrogen; and R³ is not 4-methoxyphenyl, 4-ethylphenyl or3-trifluorophenyl when R¹ is 7-chloro, and R² and R⁴ are hydrogen.

The atoms in the tricyclic system are numbered as shown in the followingformula (II).

As used in the foregoing definitions and hereinafter halo definesfluoro, chloro, bromo and iodo; C₁₋₄alkyl as a group or part of a groupencompasses the straight and branched chain saturated hydrocarbonradicals having from 1 to 4 carbon atoms such as, for example, methyl,ethyl, propyl, butyl and the like; C₁₋₆alkyl as a group or part of agroup encompasses the straight and branched chain saturated hydrocarbonradicals as defined in C₁₋₄alkyl as well as the higher homologuesthereof containing 5 or 6 carbon atoms such as, for example, pentyl orhexyl; haloC₁₋₆alkyl defines C₁₋₆alkyl wherein at least one hydrogenatom is replaced by a halo atom up to all hydrogen atoms being replacedby halo atoms, such as for example, fluoromethyl, difluoromethyl, ortrifluoromethyl.

For therapeutic use, salts of the compounds of formula (I) are thosewherein the counterion is pharmaceutically acceptable. However, salts ofacids which are non-pharmaceutically acceptable may also find use, forexample, in the preparation or purification of a pharmaceuticallyacceptable compound. All salts, whether pharmaceutically acceptable ornot are included within the ambit of the present invention.

The pharmaceutically acceptable addition salts as mentioned hereinaboveare meant to comprise the therapeutically active non-toxic acid additionsalt forms which the compounds of formula (I) are able to form. Thelatter can conveniently be obtained by treating the base form with suchappropriate acids as inorganic acids, for example, hydrohalic acids,e.g. hydrochloric, hydrobromic and the like; sulfuric acid; nitric acid;phosphoric acid and the like; or organic acids, for example, acetic,propanoic, hydroxyacetic, 2-hydroxypropanoic, 2-oxopropanoic, oxalic,malonic, succinic, maleic, fumaric, malic, tartaric,2-hydroxy-1,2,3-propanetricarboxylic, methanesulfonic, ethanesulfonic,benzenesulfonic, 4-methylbenzenesulfonic, cyclohexanesulfamic,2-hydroxybenzoic, 4-amino-2-hydroxybenzoic and the like acids.Conversely the salt form can be converted by treatment with alkali intothe free base form.

The term addition salt also comprises the hydrates and solvent additionforms which the compounds of formula (I) are able to form. Examples ofsuch forms are e.g. hydrates, alcoholates and the like.

The term “quaternary amine” as used hereinbefore defines the quaternaryammonium salts which the compounds of formula (I) are able to form byreaction between a basic nitrogen of a compound of formula (I) and anappropriate quaternizing agent, such as, for example, an optionallysubstituted alkylhalide, arylhalide or arylalkylhalide, e.g.methyliodide or benzyliodide. Other reactants with good leaving groupsmay also be used, such as alkyl trifluoromethanesulfonates, alkylmethanesulfonates, and alkyl p-toluenesulfonates. A quaternary amine hasa positively charged nitrogen. Pharmaceutically acceptable counterionsinclude chloro, bromo, iodo, trifluoroacetate and acetate. Thecounterion of choice can be made using ion exchange resin columns.

The term “stereochemically isomeric forms” as used hereinbefore definesall the possible stereoisomeric forms in which the compounds of formula(I) exist, thus, also including all enantiomers, enantiomeric mixturesand diastereomeric mixtures. Unless otherwise mentioned or indicated,the chemical designation of compounds denotes the mixture of allpossible stereoisomeric forms, said mixtures containing alldiastereomers and enantiomers of the basic molecular structure. The sameapplies to the intermediates as described herein, used to prepare endproducts of formula (I).

Pure stereoisomeric forms of the compounds and intermediates asmentioned herein are defined as isomers substantially free of otherenantiomeric or diastereomeric forms of the same basic molecularstructure of said compounds or intermediates. In particular, the term‘stereoisomerically pure’ being equivalent to ‘chirally pure’ concernscompounds or intermediates having a stereoisomeric excess of at least80% (i.e. minimum 90% of one isomer and maximum 10% of the otherpossible isomers) up to a stereoisomeric excess of 100% (i.e. 100% ofone isomer and none of the other), more in particular, compounds orintermediates having a stereoisomeric excess of 90% up to 100%, evenmore in particular having a stereoisomeric excess of 94% up to 100% andmost in particular having a stereoisomeric excess of 97% up to 100%. Theterms ‘enantiomerically pure’ and ‘diastereomerically pure’ should beunderstood in a similar way, but then having regard to the enantiomericexcess, respectively the diastereomeric excess of the mixture inquestion.

The compounds of formula (I), wherein R⁴ is hydrogen, all contain atleast 1 asymmetric center which may have the R- or S-configuration. Asused herein, the stereochemical descriptors denoting the stereochemicalconfiguration of the asymmetric center are in accordance with ChemicalAbstracts nomenclature.

Of some compounds of formula (I) and of intermediates used in theirpreparation, the absolute stereochemical configuration was notexperimentally determined. In those cases the stereoisomeric form whichwas first isolated is designated as “A” and the second as “B”, withoutfurther reference to the actual stereochemical configuration. However,said “A” and “B” stereoisomeric forms can be unambiguously characterizedby for instance their optical rotation in case “A” and “B” have anenantiomeric relationship. A person skilled in the art is able todetermine the absolute configuration of such compounds using art-knownmethods such as, for example, X-ray diffraction.

The N-oxide forms of the present compounds are meant to comprise thecompounds of formula (I) wherein one or several nitrogen atoms areoxidized to the so-called N-oxide.

Whenever used hereinafter, the term “compounds of formula (I)” is meantto include N-oxide forms, salts, quaternary amines and stereochemicallyisomeric forms. Of special interest are those compounds of formula (I)which are stereochemically pure.

Also of interest are compounds wherein

-   R¹ represents hydrogen, C₁₋₆alkyl or halo;-   R² represents hydrogen or C₁₋₆alkyl;-   R³ represents phenyl substituted with halo, cyano, C₁₋₄alkyloxy,    C₁₋₆alkyl, haloC₁₋₆alkyl; 2-thienyl or 3-thienyl, and-   R⁴ represents hydrogen; or R² and R⁴ form an extra bond.

An interesting group of compounds are those compounds of formula (I)wherein

-   R¹ represents hydrogen or halo;-   R² represents hydrogen or C₁₋₆alkyl;-   R³ represents phenyl substituted with C₁₋₆alkyl;-   R⁴ represents hydrogen; or R² and R⁴ form an extra bond.

Especially interesting are those compounds of formula (I) wherein

-   R¹ represents hydrogen, 7-chloro, 7-fluoro or 9-chloro;-   R² represents hydrogen; and-   R³ represents phenyl substituted with C₁₋₆alkyl.-   R⁴ represents hydrogen; or R² and R⁴ form an extra bond.

Most interesting are compounds (2), (21), (22), (23) and (24).

The compounds of the present invention can be prepared according toreaction scheme 1.

The compounds of formula (I) wherein R² represents C₁₋₆alkyl, saidcompounds being represented by formula (I-a) wherein R^(2a) isC₁₋₆alkyl, can be prepared from the compounds represented by formula(I-b), following art-known N-alkylation and reductive aminationreactions.

The N-alkylation reactions are conducted by reacting a compound offormula (I-b) with an alkylating agent R^(2a)—W (III) wherein W is aleaving group such as halo, e.g. chloro, bromo or iodo, or a sulfonyloxygroup, e.g. methanesulfonyloxy (mesylate) or 4-methylbenzenesulfonyloxy(tosylate) in an appropriate solvent such as an alkanol, e.g. methanol,ethanol, isopropanol; a ketone, e.g. acetone or methylisopropylketone; adipolar aprotic solvent, e.g. N,N-dimethylformamide,N,N-dimethylacetamide; in the presence of a base such as an alkali metalhydroxide or carbonate, e.g. sodium or potassium hydroxide, or sodium orpotassium carbonate. The N-alkylation reaction can also be conducted byreacting a compound of formula (I-b) with a dialkylsulphate, e.g.dimethylsulphate in water or a mixture of water and an alkanol in thepresence of a base such as sodium bicarbonate. The reaction rate can beenhanced by stirring and heating the reaction mixture, and—ifrequired—by catalyzing the N-alkylation reaction with an appropriatecatalyst such as potassium iodide.

Reductive amination reactions can be conducted by reacting a compound offormula (I-a) with an aldehyde or ketone of formula R^(2-b)═O (IV)wherein R^(2-b) represents a C₁₋₆alkanediyl radical and ═O represents anoxo-group, in an appropriate solvent such as an alkanol, e.g. methanol,ethanol, isopropanol; a dipolar aprotic solvent, e.g.N,N-dimethylformamide, N,N-dimethylacetamide; in the presence of areducing agent such as sodiumborohydride, or in the presence of hydrogenand a catalyst such as palladium. The reaction rate can be enhanced bystirring and heating the reaction mixture.

The compounds of formula (I) wherein R² and R⁴ together form an extrabond, said compounds being represented by formula (I-c), can be preparedfrom the compounds represented by the formula (I-b), following art-knownamine to imine oxidation reactions. These oxidation reactions may beconducted by reacting a compound of formula (I-b) with an oxidant suchas, for example, lead tetraacetate or manganese dioxide, in a reactioninert solvent such as a halogenated hydrocarbon e.g. dichloromethane ortrichloromethane. The reaction rate can be enhanced by stirring andoptionally heating the reaction mixture.

The compounds of formula (I-b) can be prepared from a1-(2-aminomethylphenyl)-pyrrole (V) by converting it in a salt (VI) byreaction with an acid H⁺X⁻ (VII), and reacting said salt (VI) with analdehyde of formula (VIII) in an appropriate solvent such as an alkanol,e.g. methanol, ethanol, isopropanol, at an elevated temperature,preferably at reflux temperature.

Alternatively, the intermediate of formula (V) may be reacted first withthe aldehyde R³—CHO (VIII) and the thus formed intermediate imine may becyclized in the presence of an acid H⁺X³¹ (VII) to a compound of formula(I-b).

Yet another alternative comprises N-alkylating the intermediate offormula (V) with a reagent R^(2a)—W (III) or R^(2b)═O (IV) under theconditions described before to yield an amine of formula (IX) which isfurther converted to a salt form (X) and reacted with an aldehyde (VIII)as described before.

The intermediate of formula (V) is prepared by reducing a1-(2-cyanophenyl)pyrrole (XI). Four different procedures may be used toreduce the nitrile function.

-   1. LiAlH₄/THF [S. Raines, S. Y. Chai and F. P. Palopoli; J.    Heterocyclic Chem., 13, 711-716 (1976)]-   2. i. sodium bis(2-methoxyethoxy)aluminate (Red-A1®) 70% w/w    Toluene, RT:    -   ii. NaOH 10%, RT [G. W. H. Cheeseman and S. G. Greenberg; J.        Heterocyclic Chem., 16, 241-244 (1979)]-   3. i. KBH₄/CF₃COOH, THF; ii. H₂O; iii. HCl [P. Trinka, P. Slégel    and J. Reiter; J. Prakt. Chem., 338, 675-678 (1996)]-   4. RaNi/H₂

The intermediate of formula (XI) in turn, is prepared by treating a2-aminobenzonitrile (XII) with 2,5-dimethoxyfuran in an inert solventsuch as dioxane, tetrahydrofuran in the presence of an acid such as4-chloropyridine hydrochloride, or in an acid solvent such as glacialacetic acid, at an elevated temperature, preferably at refluxtemperature.

In all these preparations, the reaction products may be isolated fromthe reaction medium and, if necessary, further purified according tomethodologies generally known in the art such as, for example,extraction, crystallization, trituration and chromatography. Inparticular, stereoisomers can be isolated chromatographically using achiral stationary phase such as, for example, Chiralpak AD (amylose 3,5dimethylphenyl carbamate) or Chiralpak AS, both purchased from DaicelChemical Industries, Ltd, in Japan.

The compounds of formula (I) may also be converted to the correspondingN-oxide forms following art-known procedures for converting a trivalentnitrogen into its N-oxide form. Said N-oxidation reaction may generallybe carried out by reacting the starting material of formula (I) with anappropriate organic or inorganic peroxide. Appropriate inorganicperoxides comprise, for example, hydrogen peroxide, alkali metal orearth alkaline metal peroxides, e.g. sodium peroxide, potassiumperoxide; appropriate organic peroxides may comprise peroxy acids suchas, for example, benzenecarboperoxoic acid or halo substitutedbenzenecarboperoxoic acid, e.g. 3-chlorobenzenecarboperoxoic acid,peroxoalkanoic acids, e.g. peroxoacetic acid, alkylhydroperoxides, e.g.tert-butyl hydroperoxide. Suitable solvents are, for example, water,lower alkanols, e.g. ethanol and the like, hydrocarbons, e.g. toluene,ketones, e.g. 2-butanone, halogenated hydrocarbons, e.g.dichloromethane, and mixtures of such solvents.

Pure stereoisomeric forms of the compounds and the intermediates of thisinvention may be obtained by the application of art-known procedures.Diastereomers may be separated by physical separation methods such asselective crystallization and chromatographic techniques, e.g. liquidchromatography using chiral stationary phases. Enantiomers may beseparated from each other by the selective crystallization of theirdiastereomeric salts with optically active acids. Alternatively,enantiomers may be separated by chromato-graphic techniques using chiralstationary phases. Said pure stereoisomeric forms may also be derivedfrom the corresponding pure stereoisomeric forms of the appropriatestarting materials, provided that the reaction occurs stereoselectivelyor stereospecifically. Preferably if a specific stereoisomer is desired,said compound will be synthesized by stereoselective or stereospecificmethods of preparation. These methods will advantageously employchirally pure starting materials. Stereoisomeric forms of the compoundsof formula (I) are obviously intended to be included within the scope ofthe invention.

The chirally pure forms of the compounds of formula (I) form a preferredgroup of compounds. It is therefore that the chirally pure forms of theintermediates of formula (II), (III) and (VI), their N-oxide forms,their salt forms and their quaternary amines are particularly useful inthe preparation of chirally pure compounds of formula (I). Alsoenantiomeric mixtures and diastereomeric mixtures of intermediates offormula (II), (ITT) and (VI) are useful in the preparation of compoundsof formula (I) with the corresponding configuration.

The compounds of formula (I), the salts, the quaternary amines and thestereochemically isomeric forms thereof are useful agents for combatingfungi in vivo. The present compounds are active against a wide varietyof fungi, such as Candida spp., e.g. Candida albicans, Candida glabrata,Candida krusei, Candida parapsilosis, Candida kefyr, Candida tropicalis;Aspergillus spp., e.g. Aspergillus fumigatus, Aspergillus niger,Aspergillus flavus; Cryptococcus neoformans; Sporothrix schenckii;Epidermophyton floccosum; Microsporum canis; Trichophyton spp., e.g.Trichophyton mentagrophytes, Trichophyton rubrum, Trichophytonquinckeanum; and several dematiaceous hyphomycetes.

In vitro experiments, including the determination of the fungalsusceptibility of the present compounds as described in thepharmacological example hereinafter, indicate that the compounds offormula (I) have a favourable intrinsic inhibitory capacity on fungalgrowth in for instance Trichophyton rubru. Other in vitro experimentssuch as the determination of the effects of the present compounds on thesterol synthesis in, for instance, Trichophyton rubrur, also demonstratetheir antifungal potency. Also in vivo experiments in a mouse model showthat the present compounds are potent antifungals when administeredintraperitoneally.

In view of the utility of the compounds of formula (I), there isprovided a method of treating warm-blooded animals, including humans,suffering from fungal infections. Said method comprises the systemic ortopical administration of an effective amount of a compound of formula(I), a N-oxide form, a salt, a quaternary amine or a possiblestereoisomeric form thereof, to warm-blooded animals, including humans.Hence, compounds of formula (I) are provided for use as a medicine, inparticular, the use of a compound of formula (I) in the manufacture of amedicament useful in treating fungal infections is provided.

The present invention also provides compositions for treating orpreventing fungal infections comprising a therapeutically effectiveamount of a compound of formula (I) and a pharmaceutically acceptablecarrier or diluent.

In view of their useful pharmacological properties, the subjectcompounds may be formulated into various pharmaceutical forms foradministration purposes. To prepare the pharmaceutical compositions ofthis invention, a therapeutically effective amount of a particularcompound, in base or addition salt form, as the active ingredient iscombined in intimate admixture with a pharmaceutically acceptablecarrier, which carrier may take a wide variety of forms depending on theform of preparation desired for administration. These pharmaceuticalcompositions are desirably in unitary dosage form suitable, preferably,for administration orally, rectally, topically, percutaneously,transungually or by parenteral injection. For example, in preparing thecompositions in oral dosage form, any of the usual pharmaceutical mediamay be employed, such as, for example, water, glycols, oils, alcoholsand the like in the case of oral liquid preparations such assuspensions, syrups, emulsions, elixirs and solutions: or solid carrierssuch as starches, sugars, kaolin, lubricants, binders, disintegratingagents and the like in the case of powders, pills, capsules and tablets.Because of their ease in administration, tablets and capsules representthe most advantageous oral dosage unit form, in which case solidpharmaceutical carriers are obviously employed. As appropriatecompositions for topical application there may be cited all compositionsusually employed for topically administering drugs e.g. creams, gel,dressings, shampoos, tinctures, pastes, ointments, salves, powders andthe like. In the compositions suitable for percutaneous administration,the carrier optionally comprises a penetration enhancing agent and/or asuitable wetting agent, optionally combined with suitable additives ofany nature in minor proportions, which additives do not cause asignificant deleterious effect to the skin. Said additives mayfacilitate the administration to the skin and/or may be helpful forpreparing the desired compositions. These compositions may beadministered in various ways, e.g., as a transdermal patch, as aspot-on, as an ointment.

Transungual compositions are in the form of a solution and the carrieroptionally comprises a penetration enhancing agent which favours thepenetration of the antifungal into and through the keratinized unguallayer of the nail. The solvent medium comprises water mixed with aco-solvent such as an alcohol having from 2 to 6 carbon atoms, e.g.ethanol.

For parenteral compositions, the carrier will usually comprise sterilewater, at least in large part. Injectable solutions, for example, may beprepared in which the carrier comprises saline solution, glucosesolution or a mixture of saline and glucose solution. Injectablesuspensions may also be prepared in which case appropriate liquidcarriers, suspending agents and the like may be employed. For parenteralcompositions, also other ingredients, to aid solubility for example,e.g. cyclodextrins, may be included. Appropriate cyclodextrins are α-,β-, γ-cyclodextrins or ethers and mixed ethers thereof wherein one ormore of the hydroxy groups of the anhydroglucose units of thecyclodextrin are substituted with C₁₋₆alkyl, particularly methyl, ethylor isopropyl, e.g. randomly methylated β-CD; hydroxyC₁₋₆alkyl,particularly hydroxyethyl, hydroxy-propyl or hydroxybutyl;carboxyC₁₋₆alkyl, particularly carboxymethyl or carboxy-ethyl;C₁₋₆alkylcarbonyl, particularly acetyl. Especially noteworthy ascomplexants and/or solubilizers are β-CD, randomly methylated β-CD,2,6-dimethyl-β-CD, 2-hydroxyethyl-β-CD, 2-hydroxyethyl-γ-CD,2-hydroxypropyl-γ-CD and (2-carboxymethoxy)propyl-β-CD, and inparticular 2-hydroxypropyl-β-CD (2-HP-β-CD).

The term mixed ether denotes cyclodextrin derivatives wherein at leasttwo cyclodextrin hydroxy groups are etherified with different groupssuch as, for example, hydroxy-propyl and hydroxyethyl.

The average molar substitution (M.S.) is used as a measure of theaverage number of moles of alkoxy units per mole of anhydroglucose. Theaverage substitution degree (D.S.) refers to the average number ofsubstituted hydroxyls per anhydroglucose unit. The M.S. and D.S. valuecan be determined by various analytical techniques such as nuclearmagnetic resonance (NMR), mass spectrometry (MS) and infraredspectroscopy (IR). Depending on the technique used, slightly differentvalues may be obtained for one given cyclodextrin derivative.Preferably, as measured by mass spectrometry, the M.S. ranges from 0.125to 10 and the D.S. ranges from 0.125 to 3.

Other suitable compositions for oral or rectal administration compriseparticles consisting of a solid dispersion comprising a compound offormula (I) and one or more appropriate pharmaceutically acceptablewater-soluble polymers.

The term “a solid dispersion” used hereinafter defines a system in asolid state (as opposed to a liquid or gaseous state) comprising atleast two components, in casu the compound of formula (I) and thewater-soluble polymer, wherein one component is dispersed more or lessevenly throughout the other component or components (in case additionalpharmaceutically acceptable formulating agents, generally known in theart, are included, such as plasticizers, preservatives and the like).When said dispersion of the components is such that the system ischemically and physically uniform or homogenous throughout or consistsof one phase as defined in thermo-dynamics, such a solid dispersion willbe called “a solid solution”. Solid solutions are preferred physicalsystems because the components therein are usually readily bioavailableto the organisms to which they are administered. This advantage canprobably be explained by the ease with which said solid solutions canform liquid solutions when contacted with a liquid medium such as thegastro-intestinal juices. The ease of dissolution may be attributed atleast in part to the fact that the energy required for dissolution ofthe components from a solid solution is less than that required for thedissolution of components from a crystalline or microcrystalline solidphase.

The term “a solid dispersion” also comprises dispersions which are lesshomogenous throughout than solid solutions. Such dispersions are notchemically and physically uniform throughout or comprise more than onephase. For example, the term “a solid dispersion” also relates to asystem having domains or small regions wherein amorphous,microcrystalline or crystalline compound of formula (I), or amorphous,microcrystalline or crystalline water-soluble polymer, or both, aredispersed more or less evenly in another phase comprising water-solublepolymer, or compound of formula (I), or a solid solution comprisingcompound of formula (I) and water-soluble polymer. Said domains areregions within the solid dispersion distinctively marked by somephysical feature, small in size, and evenly and randomly distributedthroughout the solid dispersion.

Various techniques exist for preparing solid dispersions includingmelt-extrusion, spray-drying and solution-evaporation.

The solution-evaporation process comprises the following steps

-   a) dissolving the compound of formula (I) and the water-soluble    polymer in an appropriate solvent, optionally at elevated    temperatures;-   b) heating the solution resulting under point a), optionally under    vacuum, until the solvent is evaporated. The solution may also be    poured onto a large surface so as to form a thin film, and    evaporating the solvent therefrom.

In the spray-drying technique, the two components are also dissolved inan appropriate solvent and the resulting solution is then sprayedthrough the nozzle of a spray dryer followed by evaporating the solventfrom the resulting droplets at elevated temperatures.

The preferred technique for preparing solid dispersions is themelt-extrusion process comprising the following steps:

-   -   a) mixing a compound of formula (I) and an appropriate        water-soluble polymer,    -   b) optionally blending additives with the thus obtained mixture,    -   c) heating and compounding the thus obtained blend until one        obtains a homogenous melt,    -   d) forcing the thus obtained melt through one or more nozzles;        and    -   e) cooling the melt till it solidifies.

The terms “melt” and “melting” should be interpreted broadly. Theseterms not only mean the alteration from a solid state to a liquid state,but can also refer to a transition to a glassy state or a rubbery state,and in which it is possible for one component of the mixture to getembedded more or less homogeneously into the other. In particular cases,one component will melt and the other component(s) will dissolve in themelt thus forming a solution, which upon cooling may form a solidsolution having advantageous dissolution properties.

After preparing the solid dispersions as described hereinabove, theobtained products can be optionally milled and sieved.

The solid dispersion product can be milled or ground to particles havinga particle size of less than 600 μm, preferably less than 400 μm andmost preferably less than 125 μm.

The particles prepared as described hereinabove can then be formulatedby conventional techniques into pharmaceutical dosage forms such astablets and capsules.

It will be appreciated that a person of skill in the art will be able tooptimize the parameters of the solid dispersion preparation techniquesdescribed above, such as the most appropriate solvent, the workingtemperature, the kind of apparatus being used, the rate of spray-drying,the throughput rate in the melt-extruder

The water-soluble polymers in the particles are polymers that have anapparent viscosity, when dissolved at 20° C. in an aqueous solution at2% (w/v), of 1 to 5000 mPa·s more preferably of 1 to 700 mPa·s, and mostpreferred of 1 to 100 mPa·s. For example, suitable water-solublepolymers include alkylcelluloses, hydroxyalkyl-celluloses, hydroxyalkylalkylcelluloses, carboxyalkylcelluloses, alkali metal salts ofcarboxyalkylcelluloses, carboxyalkylalkylcelluloses,carboxyalkylcellulose esters, starches, pectines, chitin derivates, di-,oligo- and polysaccharides such as trehalose, alginic acid or alkalimetal and ammonium salts thereof, carrageenans, galactomannans,tragacanth, agar-agar, gummi arabicum, guar gummi and xanthan gummi,polyacrylic acids and the salts thereof, polymethacrylic acids and thesalts thereof, methacrylate copolymers, polyvinylalcohol,polyvinylpyrrolidone, copolymers of polyvinylpyrrolidone with vinylacetate, combinations of polyvinylalcohol and polyvinylpyrrolidone,polyalkylene oxides and copolymers of ethylene oxide and propyleneoxide. Preferred water-soluble polymers are hydroxypropylmethylcelluloses.

Also one or more cyclodextrins can be used as water soluble polymer inthe preparation of the above-mentioned particles as is disclosed in WO97/18839. Said cyclodextrins include the pharmaceutically acceptableunsubstituted and substituted cyclodextrins known in the art, moreparticularly α, β or γ cyclodextrins or the pharmaceutically acceptablederivatives thereof.

Substituted cyclodextrins which can be used include polyethers describedin U.S. Pat. No. 3,459,731. Further substituted cyclodextrins are etherswherein the hydrogen of one or more cyclodextrin hydroxy groups isreplaced by C₁₋₆alkyl, hydroxyC₁₋₁₆alkyl, carboxy-C₁₋₁₆alkyl orC₁₋₁₆alkyloxycarbonylC₁₋₆alkyl or mixed ethers thereof. In particularsuch substituted cyclodextrins are ethers wherein the hydrogen of one ormore cyclodextrin hydroxy groups is replaced by C₁₋₃alkyl,hydroxyC₂₋₄alkyl or carboxyC₁₋₂alkyl or more in particular by methyl,ethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, carboxy-methyl orcarboxyethyl. As used hereinbefore, C₁₋₂alkyl represents straight orbranched chain saturated hydrocarbon radicals having 1 or 2 carbon atomssuch as methyl or ethyl; C₁₋₃alkyl encompasses the straight and branchedchain saturated hydrocarbon radicals as defined in C₁₋₂alkyl as well asthe higher homologue thereof containing 3 carbon atoms, such as propyl;C₂₋₄alkyl represents straight or branched chain saturated hydrocarbonradicals having from 2 to 4 carbon atoms such as ethyl, propyl, butyl,1-methyl-propyl and the like.

Of particular utility are the β-cyclodextrin ethers, e.g.dimethyl-β-cyclodextrin as described in Drugs of the Future, Vol. 9, No.8, p. 577-578 by M. Nogradi (1984) and polyethers, e.g. hydroxypropylβ-cyclodextrin and hydroxyethyl β-cyclodextrin, being examples. Such analkyl ether may be a methyl ether with a degree of substitution of about0.125 to 3, e.g. about 0.3 to 2. Such a hydroxypropyl cyclodextrin mayfor example be formed from the reaction between β-cyclodextrin anpropylene oxide and may have a MS value of about 0.125 to 10, e.g. about0.3 to 3. Another suitable type of substituted cyclodextrins issulfobutylcyclodextrines.

The ratio of active ingredient over cyclodextrin may vary widely. Forexample ratios of 1/100 to 100/1 may be applied. Interesting ratios ofactive ingredient over cyclodextrin range from about 1/10 to 10/1. Moreinteresting ratios of active ingredient over cyclodextrin range fromabout 1/5 to 5/1.

It may further be convenient to formulate the present benzodiazepineantifungals in the form of

nanoparticles which have a surface modifier adsorbed on the surfacethereof in an amount sufficient to maintain an effective averageparticle size of less than 1000 nm. Useful surface modifiers arebelieved to include those which physically adhere to the surface of theantifungal agent but do not chemically bond to the antifungal agent.

Suitable surface modifiers can preferably be selected from known organicand inorganic pharmaceutical excipients. Such excipients include variouspolymers, low molecular weight oligomers, natural products andsurfactants. Preferred surface modifiers include nonionic and anionicsurfactants.

Yet another interesting way of formulating the present compoundsinvolves a pharmaceutical composition whereby the present antifungalsare incorporated in hydrophilic polymers and applying this mixture as acoat film over many small beads, thus yielding a composition which canconveniently be manufactured and which is suitable for preparingpharmaceutical dosage forms for oral administration.

Said beads comprise a central, rounded or spherical core, a coating filmof a hydrophilic polymer and an antifungal agent and a seal-coatinglayer.

Materials suitable for use as cores in the beads are manifold, providedthat said materials are pharmaceutically acceptable and have appropriatedimensions and firmness. Examples of such materials are polymers,inorganic substances, organic substances, and saccharides andderivatives thereof.

The pharmaceutical compositions mentioned above may also contain afungicidally effective amount of other antifungal compounds such as cellwall active compounds. The term “cell wall active compound”, as usedherein, means any compound which interferes with the fungal cell wall.Appropriate antifungal compounds for use in combination with the presentcompounds include, but are not limited to, known azoles such asfluconazole, voriconazole, itraconazole, ketoconazole, miconazole,eberconazole, ER 30346, SCH 56592, ZD-0870, UK-292663; squaleneepoxidase inhibitors such as terbinafine and butenafine; polyenes suchas amphotericin B, nystatin or liposomal and lipid forms thereof, suchas Abelcet, AmBisome and Amphocil; purine or pyrimidine nucleotideinhibitors such as flucytosine; polyoxins and nikkomycins, in particularnikkomycin Z or nikkomycin K and others which are described in U.S. Pat.No. 5,006,513 or other chitin inhibitors; elongation factor inhibitorssuch as sordarin and analogs thereof, mannan inhibitors such aspredamycin; bactericidal/permeability-inducing (BPI) protein productssuch as XMP.97 or XMP.127; complex carbohydrate antifungal agents suchas CAN-296; (1,3)-β-glucan synthase inhibitors including papulacandins,aculeacins, echinocandins (e.g. caspofungin and micafungin); orprotegrins such as IB-367.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in unit dosage form for ease ofadministration and uniformity of dosage. Unit dosage form as used in thespecification and claims herein refers to physically discrete unitssuitable as unitary dosages, each unit containing a predeterminedquantity of active ingredient calculated to produce the desiredtherapeutic effect in association with the required pharmaceuticalcarrier. Examples of such unit dosage forms are tablets (includingscored or coated tablets), capsules, pills, suppositories, powderpackets, wafers, injectable solutions or suspensions, teaspoonfuls,tablespoonfuls and the like, and segregated multiples thereof.

Those of skill in treating warm-blooded animals suffering from diseasescaused by fungi could easily determine the therapeutically effectivedaily amount from the test results given herein. In general, it iscontemplated that a therapeutically effective daily amount would be from0.05 mg/kg to 20 mg/kg body weight.

EXPERIMENTAL PART

A. Preparation of the Intermediates

Example A.1 General Procedure for 1-(2-cyanophenyl)pyrrole

A mixture of 2-aminobenzonitrile (0.01 mole),2,5-dimethoxytetrahydrofuran (0.01 mole) and glacial acetic acid (10 ml)was heated under reflux for 30 minutes (TLC check). The acetic acid andethyl acetate formed was evaporated off and the residue was purifiedusing flash chromatography on silica gel (eluent: CH₂Cl₂). The purefraction was recrystallized from the appropriate solvent (mostlyethanol).

Example A.2 General Procedure for 1-(2-aminomethylphenyl)pyrrole

Method A: Lithium Aluminiumhydride Reduction

To a well stirred suspension of (0.05 mol) of lithium aluminum hydridein 100 ml of anhydrous tetrahydrofuran, (0.043 mole) of1-(2-cyanophenyl)pyrrole were added by small portions. Then the mixturewas maintained with stirring at room temperature for 18 hours. Additionof aqueous ethanol to destroy the excess lithium aluminium hydride,followed by filtration afforded a solution, which was then evaporated invacuo. The residue was extracted with ethyl ether, the organic layerwashed with water and dried over anhydrous sodium sulphate. Removal ofthe solvent under reduced pressure gave a crude product, which waspurified by passing through an alumina column eluting with chloroform togive 1-(2-aminomethylphenyl)pyrrole.

Method B: Red-A1® Reduction

To a stirred solution of sodium dihydrobis(2-methoxyethoxy)aluminate(100 ml of 70% w/w solution in toluene) in dry toluene (100 ml) wasadded dropwise, over one hour, a solution of 1-(2′-cyanophenyl)pyrrole(16.8 g, 0.1 mole) in dry toluene. After 1.5 hours, aqueous sodiumhydroxide (10%, 100 ml) was cautiously added. The aqueous layer wasseparated off and extracted with toluene. The combined organic layerswere washed with saturated sodium chloride solution, dried (magnesiumsulfate) and evaporated, and the residue vacuum-distilled. The productwas converted in its hydrochloride by treatment with iPrOH/HCl solution.

Method C: KBH₄/CF₃COOH Reduction

To a mixture of potassium borohydride (0.55 g, 0.01 mol) and THF (10 ml)a solution of trifluoroacetic acid (1.1 g, 0.01 mol) in THF (1 ml) wasadded dropwise at a rate so as to keep the reaction temperature between15-20° C. To the reduction mixture thus obtained a solution of1-(3-chloro-2-cyanophenyl)pyrrole (0.6 g, 0.003 mol) in THF (15 ml) wasadded at a rate so as to keep the reaction temperature between 25-30° C.The reaction mixture thus obtained was stirred for further 6 hours atroom temperature. Then water (100 ml) was added and the mixture wasextracted with CH₂Cl₂ (3×), the organic layer washed with H₂O (2×), NaCl(1×), dried (MgSO₄), filtered and evaporated.

The residue was converted to its hydrochloride by treatment withiPrOH/HCl (3 ml) solution. Recrystallisation from iPrOH gave crystals.

Method D: Raney-Nickel Reduction

A mixture of 1-(3-chloro-2-cyanophenyl)pyrrole (16.3 g, 0.08 mol), RaNi(2 g), thiophene (2 ml) in methanol saturated with ammonia (250 ml) wascooled to circa 14° C. and hydrogenated at atmospheric pressure. Afteruptake of two equivalents of hydrogen, the catalyst was filtered off andthe solvent was evaporated. The residue was converted in itshydrochloride by treatment with iPrOH/HCl (16 ml) solution. Besides theaimed product also dehalogenated product was formed as a minor sideproduct.

Example A.3 Specific Procedure for1-(2′-aminomethyl-3′-chlorophenyl)pyrrole

A solution of 2-amino-6-chloro-benzonitrile (30 g),2,5-dimethoxytetrahydrofuran (30 g) and 4-chloropyridine hydrochloride(17 g) in dioxane was heated to 70° C. for 2.5 h. The solvent wasevaproated in vacuo. Dichloromethane was then added to the residue andthe spent 4-chloropyridine hydrochloride filtered off. Thedichloromethane was evaporated in vacuo to give1-(3′chloro-2′-cyanophenyl)pyrrole (35 g)

A solution of 1-(3′chloro-2′-cyanophenyl)pyrrole (10 g) in diethyl ethersolution was added dropwise to a solution of 1M lithium aluminiumhydride in ether solution (100 ml) under nitrogen gas. After additionwas complete, the reaction mixture was refluxed for 2 h. and thenallowed to stir at room temperature overnight (18 h). The mixture wassubsequently quenched with water (4 ml), 2N NaOH (8 ml) and water (16ml). Stirring was continued for a further 1 h after addition. Thecrystalline salts were filtered and the filtrate dried over anhydrousmagnesium sulphate. After filtering, the ether filtrate was evaporatedin vacuo to give the title compound (9.4 g) as a colourless oil.

B. Preparation of the Final Compounds

Example B1 General Procedure

A solution of (0.05 mol) 1-(2-aminomethylphenyl)pyrrole and an aldehyde(0.05 mol) in ethanol (100 ml) was heated under reflux for 4 hours. Thesolution was evaporated and the residue was dissolved in ether andethereal hydrogen chloride was added. After stirring for 30 minutes, theprecipitate was filtered off and recrystallized from an appropriatealcohol.

Example B.2 General Procedure for Alkylation of7-chloro-5,6-dihydro-4H-pyrrolo[1,2-a][1,4]benzodiazepine

A solution of the appropriate alkylhalide (1 mmole) in CH₃CN (5 ml) isadded dropwise to a solution of7-chloro-5,6-dihydro-4H-pyrrolo[1,2-a][1,4]benzodiazepine (1 mmole) andtriethylamine (1 mmole) in CH₃CN (5 ml) at room temperature. Afterstirring at room temperature for 1 to 48 hours, the mixture isevaporated in vacuo and subjected to column chromatography on silicagel.

Example B.3 Specific Procedure for4-(4′-ethylphenyl)-5-methyl-7-chlorobenzo[1,2]-pyrrolo-[1,2a][1,4]diazepine

A solution of 1-(2′-aminomethyl-3′-chlorophenyl)pyrrole (1.5 g) and4-ethylbenzaldehyde (1 g) in ethanol was refluxed 2 h. The solution wascooled to 0° C. and a solution of concentrated (45%) hydrobromic acid (4ml) in glacial acetic acid (9 ml) was added dropwise. The solution wasstirred at room temperature for a further 1 h. after which the compound(2) was filtered off (2.2 g). The compound (2) (0.5 g) was added to asaturated aqueous solution of sodium bicarbonate containing a smallquantity of methanol. A slight excess of dimethyl sulphate was added andthe reaction mixture stirred at room temperature for 30 minutes. Theresulting solid was filtered, dried and purified using a silica gelcolumn using dichloromethane as eluant to give compound (10) (0.17 g) asa white solid.

Example B.4 Procedure for Compound 21

A solution of compound (13) was stirred for three days at roomtemperature in dichloromethane in the presence of an excess of manganesedioxide. The suspension was filtered and the filtrate was purified bycolumn chromatography on silica gel yielding a viscous oil.

Example B.5 Procedure for Resolution of (2) into Compounds (24) and (29)

A mixture of 1-(2-aminomethyl-3-chlorophenyl)pyrrole hydrochloride(prepared according to A.2 method D) (0.024 mol) and 4-ethylbenzaldehyde(0.024 mol) in EtOH (50 ml) was stirred and refluxed for 2 hours. Themixture was crystallized overnight. The crystals were filtered off andwashed 3 times with EtOH (10 ml) and then dried (vacuum, 60° C.). Yield:4.7 g. The residue was purified by high performance liquidchromatography over Chiralpak AD (eluent: 100% EtOH). The productfractions were collected and the solvent was evaporated. The yield offraction A (0.968 g) and of fraction B (0.92 g) were converted into thehydrochloric acid salt (1:1) with 2-propanol and HCl (6N), filtered offand dried. Yield: 0.911 g of compound 29 (11.5%) and 0.861 g of compound24 (11%).

Following the foregoing examples, the following compounds have beenprepared.

TABLE 1

Comp. No. R¹ R² R⁴ R³ Physical data 1 7-Cl H H 4-MeO—C₆H₄— .HBr (1:1);m.p.: 242-243° C. 2 7-Cl H H 4-Et—C₆H₄— .HBr (1:1); m.p.: 244-245° C. 37-Cl H H 3-CF₃—C₆H₄— .HBr (1:1); m.p.: 226-227° C. 4 H H H 4-iPr—C₆H₄—.HBr (1:1); m.p.: 251-252° C. 5 7-Cl H H 4-Me—C₆H₄— .HBr (1:1); m.p.:228-229° C. 6 7-Cl H H 4-iPr—C₆H₄— .HBr (1:1); m.p.: 338-339° C. 7 7-ClH H 4-tBu—C₆H₄— .HBr (1:1); m.p.: 245-246° C. 8 7-Cl H H 4-nBu—C₆H₄—.HBr (1:1); m.p.: 227-228° C. 9 7-Cl H H 4-nPr—C₆H₄— .HBr (1:1); m.p.:238-239° C. 10 7-Cl CH₃ H 4-Et—C₆H₄— m.p.: 91-92° C. 11 8-Cl H H4-Et—C₆H₄— m.p.: 40-41° C. 12 7-Cl H H 3-thienyl .HBr (1:1); m.p.:237-238° C. 13 H H H 4-Et—C₆H₄— .HBr (1:1); m.p.: 259-260° C. 14 7-F H H4-iPr—C₆H₄— m.p.: 116-118° C. 15 7-F H H 4-Et—C₆H₄— m.p.: 133-134° C. 169-Cl H H 4-Et—C₆H₄ .HBr (1:1); m.p.: 243-244° C. 17 9-Cl H H 4-OEt—C₆H₄.HBr (1:1); m.p.: 248-249° C. 18 9-Cl H H 4-iPr—C₆H₄ .HBr (1:1); m.p.:251-252° C. 19 9-Cl H H 4-Me—C₆H₄ .HBr (1:1); m.p.: 244-245° C. 20 9-ClH H 4-CN—C₆H₄ .HBr (1:1); m.p.: 248-249° C. 21 H double bond 4-Et—C₆H₄oil 22 7-Cl double bond 4-Et—C₆H₄ m.p.: 208-208° C. 23 9-Cl double bond4-Et—C₆H₄ m.p.: 124-125° C. 24 7-Cl H H 4-Et—C₆H₄ .HCl (1:1);(B)-isomer; m.p.: 262° C. 25 7-Cl H H 3-Et—C₆H₄ 26 7-CH₃ H H 4-Br—C₆H₄.HBr (1:1); m.p.: 256-257° C. 27 7-CH₃ H H 4-Et—C₆H₄ .HBr (1:1); m.p.:257-258° C. 28 7-CH₃ H H 4-Cl—C₆H₄ .HBr (1:1); m.p.: 250-251° C. 29 7-ClH H 4-Et—C₆H₄ .HCl (1:1); (A)-isomer; m.p.: 250° C. 30 7-CH₃ H H4-CH₃S—C₆H₄ .HBr (1:1); m.p.: 257-258° C. 31 7-SCH₃ H H 4-Et—C₆H₄ .HBr(1:1); m.p.: 239-240° C. 32 10-Cl H H 4-Et—C₆H₄ .HBr (1:1); m.p.:249-250° C. 33 7-OMe H H 4-Et—C₆H₄ .HBr (1:1); m.p.: 254-255° C. 34 7-ClH H 2-Et—C₆H₄

C. Pharmacological Examples Example C.1 Measurement of AntifungalActivity In Vitro

The test compounds were dissolved at a concentration of 10⁻² M indimethyl sulfoxide (DMSO) and diluted into CYG broth (Odds, F.C.Antimicrobial Agents and Chemotherapy 1992; 36: 1727-1737) to give afinal concentration of 10, 3.2, 1, 0.32, 0.1, 0.03, 0.01, 0.003 and0.001 μM. For some compounds the tests were done at intermediateconcentrations. Cultures were inoculated with Candida kefyr to aninitial concentration of 10⁴/ml and with the other fungal species to anequivalent concentration determined by turbidimetry. Cultures wereincubated in the wells of microdilution plates at 37° C. for 48 h (C.kefyr) and at 30° C. for 5-7 days (T. rubrum). Growth in wellscontaining test compounds was estimated turbidimetrically as apercentage of growth in compound-free controls and the lowestconcentration of compound that inhibited the growth of an isolate below35% of control growth was recorded as the lowest active dose (LAD).

Example C2 Determination of Fungal Susceptibility

A panel single isolates of the dermatophytes Sporothrix schenckii;Microsporum canis; Trichophyton rubrum; Trichophyton mentagrophyte;Candida parapsilosis; Cryptococcus neoformans; and Aspergillus fumigatuswere used to evaluate the activity of the test compounds in vitro.Inocula were prepared as broth cultures (yeasts) or as suspensions offungal material made from agar slope cultures (moulds). The testcompounds were pipetted from dimethylsulfoxide stock solution into waterto provide a series of 10-fold dilutions. The fungal inocula weresuspended in the growth medium CYG (F.C. Odds, Journal of ClinicalMicrobiology, 29, 2735-2740, 1991) at approximately 50,000colony-forming units (CFU) per ml and added to the aqueous test drugs.

The cultures were set up in the 96 wells of plastic microdilution platesand they were incubated for 2 days at 37° C. (Candida spp.) or for 5days at 30° C. (other fungi). Growth in the microcultures was measuredby their optical density (OD) measured at a wavelength of 405 nm. The ODfor cultures with test compounds was calculated as a percentage of theOD for control cultures, i.e. the OD for cultures without testcompounds. Inhibition of growth to 35% of control or less was recordedas significant inhibition.

Minimal inhibitory concentration (MIC; in 10⁻⁶ M) of some of thecompounds of formula (I) for the tested species are listed in Table.

TABLE 2a MIC values in 10⁻⁶ M Infection Comp. 1 Comp. 2 Comp. 3Sporothrix schenkii >10 >10 >10 Microsporum canis 5.5 0.32 2.1Trichophyton rubrum 2.1 0.03 2.1 Trichophyton mentagrophytes 2.1 0.323.2 Candida parapsilosis >10 1 >10 Cryptococcus neoformans >10 >10 >10Aspergillus fumigatus 10 3.2 10

TABLE 2b MIC values in 10⁻⁶ M Infection Comp. 4 Comp. 5 Comp. 6Sporothrix schenkii >10 >10 >10 Microsporum canis 10 1 0.32 Trichophytonrubrum 3.2 0.32 0.32 Trichophyton mentagrophytes 3.2 1 1 Candidaparapsilosis >10 >10 10 Cryptococcus neoformans >10 >10 >10 Aspergillusfumigatus >10 >10 >10

TABLE 2c MIC values in 10⁻⁶ M Infection Comp. 7 Comp. 8 Comp. 9Sporothrix schenkii 10 >10 >10 Microsporum canis 0.32 1 0.32Trichophyton rubrum 0.1 0.1 0.1 Trichophyton mentagrophytes 1 1 0.32Candida parapsilosis 10 >10 3.2 Cryptococcus neoformans >10 >10 >10Aspergillus fumigatus >10 >10 >10

TABLE 2d MIC values in 10⁻⁶ M Infection Comp. 10 Comp. 11 Comp. 12Sporothrix schenkii >10 >10 >10 Microsporum canis >10 >10 >10Trichophyton rubrum 3.2 3.2 3.2 Trichophyton mentagrophytes 10 3.2 >10Candida parapsilosis >10 >10 >10 Cryptococcus neoformans >10 >10 >10Aspergillus fumigatus >10 >10 >10

TABLE 2e MIC values in 10⁻⁶ M Infection Comp. 13 Comp. 14 Comp. 15Sporothrix schenkii >10 >10 >10 Microsporum canis >10 10 3.2Trichophyton rubrum 1 0.32 0.32 Trichophyton mentagrophytes 3.2 1 1Candida parapsilosis >10 >10 10 Cryptococcus neoformans >10 >10 >10Aspergillus fumigatus >10 >10 10

TABLE 2f MIC values in 10⁻⁶ M Infection Comp. 16 Comp. 17 Comp. 18Sporothrix schenkii >10 >10 >10 Microsporum canis 1 >10 >10 Trichophytonrubrum 0.1 10 1 Trichophyton mentagrophytes 1 10 1 Candidaparapsilosis >10 >10 >10 Cryptococcus neoformans >10 >10 >10 Aspergillusfumigatus >10 >10 >10

TABLE 2g MIC values in 10⁻⁶ M Infection Comp. 19 Comp. 20 Comp. 21Sporothrix schenkii >10 >10 10 Microsporum canis 10 >10 0.32Trichophyton rubrum 0.32 0.32 0.03 Trichophyton mentagrophytes 3.2 >100.32 Candida parapsilosis >10 >10 3.2 Cryptococcusneoformans >10 >10 >10 Aspergillus fumigatus >10 >10 1

D. Composition Example

“Active ingredient” (A.I.) as used throughout these examples relates toa compound of formula (I), a N-oxide, a salt, a quaternary amine or astereochemically isomeric form thereof.

Example D1 Injectable Solution

1.8 Grams methyl 4-hydroxybenzoate and 0.2 grams sodium hydroxide weredissolved in about 0.5 l of boiling water for injection. After coolingto about 50° C. there were added while stirring 0.05 grams propyleneglycol and 4 grams of the active ingredient. The solution was cooled toroom temperature and supplemented with water for injection q.s. ad 1 l,giving a solution comprising 4 mg/ml of active ingredient. The solutionwas sterilized by filtration and filled in sterile containers.

Example D2 Transungual Composition

0.144 g KH₂PO₄, 9 g NaCl, 0.528 g Na₂HPO₄.2H₂O was added to 800 ml H₂Oand the mixture was stirred. The pH was adjusted to 7.4 with NaOH and500 mg NaN₃ was added. Ethanol (42 v/v %) was added and the pH wasadjusted to 2.3 with HCl. 15 mg active ingredient was added to 2.25 mlPBS (Phosphate Buffer Saline)/Ethanol (42%; pH 2.3) and the mixture wasstirred and treated with ultrasound. 0.25 ml PBS/Ethanol (42%; pH 2.3)was added and the mixture was further stirred and treated withultrasound until all active ingredient was dissolved, yielding thedesired transungual composition.

Example D3 Oral Drops

500 Grams of the A.I. was dissolved in 0.5 l of a sodium hydroxidesolution and 1.5 l of the polyethylene glycol at 60-80° C. After coolingto 30-40° C. there were added 35 l of polyethylene glycol and themixture was stirred well. Then there was added a solution of 1750 gramsof sodium saccharin in 2.5 l of purified water and while stirring therewere added 2.5 l of cocoa flavor and polyethylene glycol q.s. to avolume of 50 l, providing an oral drop solution comprising 10 mg/ml ofA.I. The resulting solution was filled into suitable containers.

Example D4 Capsules

20 Grams of the A.I., 6 grams sodium lauryl sulfate, 56 grams starch, 56grams lactose, 0.8 grams colloidal silicon dioxide, and 1.2 gramsmagnesium stearate were vigorously stirred together. The resultingmixture was subsequently filled into 1000 suitable hardened gelatincapsules, comprising each 20 mg of the active ingredient.

Example D5 Film-Coated Tablets

Preparation of Tablet Core

A mixture of 100 grams of the A.I., 570 grams lactose and 200 gramsstarch was mixed well and thereafter humidified with a solution of 5grams sodium dodecyl sulfate and 10 grams polyvinylpyrrolidone in about200 ml of water. The wet powder mixture was sieved, dried and sievedagain. Then there was added 100 grams micro-crystalline cellulose and 15grams hydrogenated vegetable oil. The whole was mixed well andcompressed into tablets, giving 10.000 tablets, each containing 10 mg ofthe active ingredient.

Coating

To a solution of 10 grams methyl cellulose in 75 ml of denaturatedethanol there was added a solution of 5 grams of ethyl cellulose in 150ml of dichloromethane. Then there were added 75 ml of dichloromethaneand 2.5 ml 1,2,3-propanetriol. 10 Grams of polyethylene glycol wasmolten and dissolved in 75 ml of dichloromethane. The latter solutionwas added to the former and then there were added 2.5 grams of magnesiumoctadecanoate, 5 grams of polyvinylpyrrolidone and 30 ml of concentratedcolour suspension and the whole was homogenated. The tablet cores werecoated with the thus obtained mixture in a coating apparatus.

Example D6 2% Cream

Stearyl alcohol (75 mg), cetyl alcohol (20 mg), sorbitan monostearate(20 mg) and isopropyl myristate (10 mg) are introduced in a doublewalljacketed vessel and heated until the mixture has completely molten. Thismixture is added to a seperately prepared mixture of purified water,propylene glycol (200 mg) and polysorbate 60 (15 mg) having atemperature of 70 to 75° C. while using a homogenizer for liquids. Theresulting mixture is allowed to cool to below 25° C. while continuouslymixing. A solution of A.I. (20 mg), polysorbate 80 (1 mg) and purifiedwater q.s. ad 1 g and a solution of sodium sulfite anhydrous (2 mg) inpurified water are next added to the emulsion while continuously mixing.The cream is homogenized and filled into suitable tubes.

Example D7 2% Cream

A mixture of A.I. (2 g), phosphatidyl choline (20 g), cholesterol (5 g)and ethyl alcohol (10 g) is stirred and heated at 55-60° C. untilcomplete solution and is added to a solution of methyl paraben (0.2 g),propyl paraben (0.02 g), disodium edetate (0.15 g) and sodium chloride(0.3 g) in purified water (ad 100 g) while homogenizing.Hydroxypropylmethylcellulose (1.5 g) in purified water is added and themixing is continued until swelling is complete.

1. A compound having the formula

a N-oxide form, a salt, a quaternary amine or a stereochemicallyisomeric form thereof, wherein R¹ is selected from the group consistingof hydrogen, C₁₋₆alkyl, C₁₋₄alkylthio, C₁-₄alkyloxy, and halo; R² isselected from the group consisting of hydrogen and C₁₋₆alkyl; R³ isselected from the group consisting of phenyl and 3-thienyl, wherein thephenyl is substituted with halo, cyano, C₁₋₄alkyloxy, C₁₋₄alkylthio,C₁₋₆alkyl, or haloC₁₋₆alkyl; and R⁴ is hydrogen; or R² and R⁴ form anextra bond; provided that R³ is not 4-methylphenyl, 4-methoxyphenyl or4-isopropylphenyl when R¹, R² and R⁴ are hydrogen; R³ is not4-methoxyphenyl, 4-ethylphenyl or 3-trifluorophenyl when R¹ is 7-chloro,and R² and R⁴ are hydrogen; and R³ is not 4-chlorophenyl when R¹ ishydrogen and R² and R⁴ form an extra bond.
 2. The compound as in claim 1wherein R¹ is selected from the group consisting of hydrogen, C₁₋₆alkyland halo; R² is selected from the group consisting of hydrogen andC₁₋₆alkyl; R³ is selected from the group consisting of phenyl and3-thienyl, wherein the phenyl is substituted with halo, cyano,C₁₋₄alkyloxy, C₁₋₆alkyl, or haloC₁₋₆alkyl; and R⁴ is hydrogen; or R² andR⁴ form an extra bond.
 3. The compound as in claim 1 wherein R¹ isselected from the group consisting of hydrogen and halo; R² is selectedfrom the group consisting of hydrogen and C₁₋₆alkyl; R³ is phenylsubstituted with C₁₋₆alkyl; and R⁴ is hydrogen; or R² and R⁴ form anextra bond.
 4. The compound as in claim 1 wherein R¹ is selected fromthe group consisting of hydrogen, 7-chloro, 7-fluoro or 9-chloro; R² ishydrogen; R³ is phenyl substituted with C₁₋₆alkyl; and R⁴ is hydrogen;or R² and R⁴ form an extra bond.
 5. The compound as in claim 1 whereinR³ is selected from the group consisting of phenyl substituted withhalo, cyano, C₁₋₄alkyloxy, C₁₋₄alkylthio, C₁₋₆alkyl or haloC₁₋₆alkyl. 6.The compound as in claim 1 wherein R³ is selected from the groupconsisting of phenyl substituted with C₁₋₆alkyl.
 7. The compound as inclaim 6 wherein R³ is phenyl substituted with ethyl.
 8. The compound asin any of claim 5, 6 or 7 wherein the phenyl representing R³ issubstituted in 4-position.
 9. The compound as in any of claim 5, 6 or 7wherein the phenyl representing R³ is substituted in 3-position.
 10. Thecompound as in any one of claims 1-7 wherein R² and R⁴ form an extrabond.
 11. The compound as in claim 8 wherein R² and R⁴ form an extrabond.
 12. The compound as in claim 9 wherein R² and R⁴ form an extrabond.
 13. The compound as in claim 10 wherein R¹ is hydrogen.
 14. Thecompound as in claim 11 wherein R¹ is hydrogen.
 15. The compound as inclaim 12 wherein R¹ is hydrogen.
 16. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier, and as activeingredient a therapeutically effective amount of a compound as inclaim
 1. 17. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier, and as active ingredient a therapeutically effectiveamount of a compound as in claim
 8. 18. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier, and as activeingredient a therapeutically effective amount of a compound as in claim9.
 19. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier, and as active ingredient a therapeutically effectiveamount of a compound as in claim
 10. 20. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier, and as activeingredient a therapeutically effective amount of a compound as in claim11.
 21. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier, and as active ingredient a therapeutically effectiveamount of a compound as in claim
 12. 22. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier, and as activeingredient a therapeutically effective amount of a compound as in claim13.
 23. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier, and as active ingredient a therapeutically effectiveamount of a compound as in claim
 14. 24. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier, and as activeingredient a therapeutically effective amount of a compound as in claim15.